Type ii restriction endonuclease, pme i, obtainable from pseudomonas mendocina and a process for producing the same

Abstract

The present invention provides a novel type II restriction endonuclease obtainable from Pseudomonas mendocina. The endonuclease known as Pme I, recognizes the following nucleotide sequence and has a cleavage point indicated by the arrows: 5'-G T T T↓A A A C-3' 3'-C A A A↑T T T G-5' Also described is a process for obtaining Pme I from Pseudomonas mendocina.

What is claimed is: 
     
       1. A substantially pure Type II restriction endonuclease obtainable from Pseudomonas mendocina (ATCC#55181) recognizing the following base sequence in double-stranded deoxyribonucleic acid molecules: 5'-G T T T↓A A A C-3'   3'-C A A A↓T T T G-5' and having a cleavage position defined by the arrows.     
     
     
       2. The type II restriction endonuclease of claim 1, cleaving double-stranded deoxyribonucleic acid lambda cI857 in two and adeno-2 in one position. 
     
     
       3. A method for obtaining the Type II restriction endonuclease of claim 1, comprising cultivating a sample of Pseudomonas mendocina under conditions favoring the production of said endonuclease and separating said endonuclease therefrom. 
     
     
       4. The type II restriction endonuclease of claim 1, wherein the restriction endonuclease is inactivated by incubation at 65° C. for 20 minutes. 
     
     
       5. The type II restriction endonuclease of claim 1, wherein the restriction endonuclease is purified from Pseudomonas mendocina(ATCC#55181).